Method utilizing RNA interference technique to breed corn with enhanced content of amylose

A technology for amylose content and corn, which is applied in the direction of recombinant DNA technology, DNA/RNA fragments, botanical equipment and methods, etc., to achieve the effects of increasing content, speeding up the cultivation of production varieties, and reducing the amount of synthesis

Inactive Publication Date: 2009-12-02
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Application Information

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Problems solved by technology

However, conventional cross-breeding techniques have also exposed

Method used

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  • Method utilizing RNA interference technique to breed corn with enhanced content of amylose
  • Method utilizing RNA interference technique to breed corn with enhanced content of amylose
  • Method utilizing RNA interference technique to breed corn with enhanced content of amylose

Examples

Experimental program
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Embodiment 1

[0018] Example 1. Increase the amylose content in corn by introducing RNAi vector of sbe IIb gene into corn

[0019] Maize inbred lines 178, 478, 501, Huang C and ZP501K are universal inbred line materials, which come from the National Germplasm Bank. Restriction endonuclease, T 4 DNA ligase was purchased from NEB Company (UK), pGEM-T easy vector was purchased from Promega Company (USA), Tag DNA polymerase was purchased from TaKaRa Company (Japan), E.coli strain DH5α was purchased from Beijing Tiangen the company.

[0020] 1. Construction of RNAi vector pBAC413 of sbeIIb gene

[0021] 1. Cloning of starch branching enzyme gene sbeIIb

[0022] After inputting the mRNA sequence of sbe IIb (AF072725) in the network database (http: / / jura.wi.mit.edu / bioc / siRNAext / ), a series of optimal siRNA sites were obtained, and DNAMAN was used for Not I, HindIII , EcoRI, BamHI and other commonly used restriction sites analysis. According to the above steps, a total of 1162 bp of sbeIIb mRNA sequen...

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Abstract

The invention discloses a method for breeding corn with the enhanced content of amylase. The method for breeding corn with the enhanced content of amylase comprises the following steps that: a recombinant carrier which expresses the following interference RNA molecules is introduced into corn, and corn with silenced starch branching enzyme SBEIIb coding genes is obtained, i.e. the corn with the enhanced content of amylase; and one chain sequence of the interference RNA molecules is shown in a sequence 1 in a sequence table, and the other chain sequence is shown in a sequence 2 in the sequence table. An experimental result shows that the content of the amylase in the corn can be enhanced by 15% through the method. In addition, the method can overcome the limitations of a traditional breeding method, can rapidly obtain a target character by a gene engineering technique and can be combined with conventional breeding so as to quicken the breeding of the production variety of high-amylose corn starch.

Description

Technical field [0001] The invention relates to a method for cultivating corn with increased amylose content, in particular to a method for cultivating corn with increased amylose content by using RNA interference technology. Background technique [0002] The development of the corn industry has important strategic significance to my country's agricultural economy. The development of high-straight-chain corn can extend the intensive and deep processing industrial chain of corn, which is of far-reaching significance to increase the added value of corn planting and increase farmers' yield and income; at the same time, it also plays a positive role in promoting the development, application and promotion of new degradable environmentally friendly materials. [0003] The starch in corn accounts for about 70%-75% of the dry weight of ordinary corn kernels, of which amylopectin accounts for about 75% of the total starch content, while amylose accounts for only about 25%. With the rapid ...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/82A01H1/00A01H5/00C12N15/113
Inventor 张晓东陈绪清杨凤萍张立全梁荣奇郭新梅
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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